Plant Molecular Biology

, Volume 23, Issue 6, pp 1151–1164 | Cite as

Organization and structure of the 1-aminocyclopropane-1-carboxylate oxidase gene family from Petunia hybrida

  • Xiaoyan Tang
  • Hong Wang
  • Amanda S. Brandt
  • William R. Woodson
Research Article


In this paper we present the structural analysis of the 1-aminocyclopropane-1-carboxylate (ACC) oxidase gene family from Petunia hybrida. Southern blot analysis and restriction endonuclease mapping showed that two cloned regions of the petunia genome contained sequences highly homologous to a previously isolated ACC oxidase cDNA clone. Nucleotide sequencing of these two regions of the genome showed that each contained two tandemly arranged genes designated ACO1, ACO2, ACO3 and ACO4. Comparison of the nucleotide sequences of the cloned genomic regions with the cDNA clone pPHEFE indicated that ACO1 encoded the transcript in 4 exons interrupted by 3 introns. The other three members of the petunia ACC oxidase gene family shared identical intron numbers and positions with ACO1 and their exons were greater than 80% homologous. Nucleotide substitutions and deletions in the ACO2 gene indicate that it likely represents a pseudogene. Overall homology between ACO1 and ACO2 indicates that this gene cluster arose by a more recent duplication event than the gene duplication giving rise to the ACO3 and ACO4 cluster. The 5-flanking sequences share little overall homology between members of this gene family. However, sequences which likely make up the core promoter of these genes including the TATA box are highly homologous. RNA-based PCR amplification of ACC oxidase cDNAs from ethylene-treated corollas and wounded leaves revealed transcripts for ACO1, ACO3 and ACO4 indicating that at least three of these genes are transcriptionally active. The proteins encoded by ACO1, ACO3 and ACO4 share more than 90% identity with one another and more than 70% identity with ACC oxidases from other species. The ACC oxidase proteins share significant sequence homology with other enzymes that require Fe(II) and ascorbate for catalytic activity.

Key words

1-aminocyclopropane-1-carboxylate oxidase ethylene biosynthesis gene cluster gene duplication 


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  1. 1.
    Balaqué, C, Watson, CF, Turner, AJ, Rouge, P, Picton, S, Pech, J-C, Grierson, D: Isolation of a ripening and wound-induced cDNA from Cucumis melo L. encoding a protein with homology to the ethylene-forming enzyme. Eur J Biochem 212: 27–34 (1993).Google Scholar
  2. 2.
    Boller, T: Ethylene in pathogenesis and disease resistance. In: Mattoo, AK, Suttle, JC (eds) The Plant Hormone Ethylene, pp. 293–314. CRC Press, Boca Raton, FL (1991).Google Scholar
  3. 3.
    Borochov, A, Woodson, WR: Physiology and biochemistry of flower petal senescence. Hort Rev 11: 15–43 (1989).Google Scholar
  4. 4.
    Botella, JR, Schlagnhaufer, CD, Arteca, JM, Arteca, RN, Phillips, AT: Identification of two new members of the 1-aminocyclopropane-1-carboxylate synthase-encoding multigene family in mung bean. Gene 123: 249–253 (1993).Google Scholar
  5. 5.
    Bouzayen, M, Cooper, W, Barry, C, Zegzouti, H, Hamilton, AJ, Grierson, D: EFE multigene family in tomato plants: expression and characterization. In: Pech, JC, Latché, A, Balagué, C (eds) Cellular and Molecular Aspects of the Plant Hormone Ethylene, pp. 76–81. Kluwer Academic Publishers, Dordrecht, Netherlands (1993).Google Scholar
  6. 6.
    Brady, CJ, Speirs, J: Ethylene in fruit ontogeny and abscission. In: Mattoo, AK, Suttle, JC (eds) The Plant Hormone Ethylene, pp. 235–258. CRC Press, Boca Raton, FL (1991).Google Scholar
  7. 7.
    Britsch, L, Ruhnau-Brich, B, Forkmann, G: Molecular cloning, sequence analysis, and in vitro expression of flavanone 3β-hydroxylase from Petunia hybrida. J Biol Chem 267: 5380–5387 (1992).Google Scholar
  8. 8.
    Broglie, KE, Biddle, P, Cressman, R, Broglie, R: Functional analysis of DNA sequences responsible for ethylene regulation of a bean chitinase gene in transgenic tobacco. Plant Cell 1: 599–607 (1989).Google Scholar
  9. 9.
    Brown, JWS: A catalogue of splice junctions and putative branch point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).Google Scholar
  10. 10.
    Deikman, J, Kline, R, Fischer, RL: Organization of ripening and ethylene regulatory regions in a fruit-specific promoter from tomato (Lycopersicon esculentum). Plant Physiol 100: 2013–2017 (1992).Google Scholar
  11. 11.
    Dellaporta, SL, Wood, J, Hicks, JB: A plant DNA miniprep: Version II. Plant Mol Biol Rep 1: 19–21 (1983).Google Scholar
  12. 12.
    Devereux, P, Haeberli, P, Smithies, O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acid Res 12: 387–395 (1984).Google Scholar
  13. 13.
    Dong, JG, Kim, WT, Yip, WK, Thompson, GA, Li, L, Bennett, AB, Yang, SF: Cloning of a cDNA encoding 1-aminocyclopropane-1-carboxylate synthase and expression of its mRNA in ripening apple fruit. Planta 185: 38–45 (1991).Google Scholar
  14. 14.
    Dong, JG, Olson, D, Silverstone, A, Yang, SF: Sequence of a cDNA coding for a 1-aminocyclopropane-1-carboxylate oxidase homolog from apple fruit. Plant Physiol 98: 1530–1531 (1992).Google Scholar
  15. 15.
    Dong, JG, Fernández-Maculet, JC, Yang, SF: Purification and characterization of 1-aminocyclopropane-1-carboxylate oxidase from apple fruit. Proc Natl Acad Sci USA 89: 9789–9793 (1992).Google Scholar
  16. 16.
    Felix, G, Grosskopf, DG, Regenass, M, Basse, CW, Boller, T: Elicitor-induced ethylene biosynthesis in tomato cells. Characterization and use as a bioassay for elicitor action. Plant Physiol 97: 19–25 (1991).Google Scholar
  17. 17.
    Guy, M, Kende, H: Conversion of 1-aminocyclopropane-1-carboxylic acid to ethylene by isolated vacuoles of Pisum sativum L. Planta 160: 281–287 (1984).Google Scholar
  18. 18.
    Hamilton, AJ, Lycett, GW, Grierson, D: Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346: 284–287 (1990).Google Scholar
  19. 19.
    Hamilton, AJ, Bouzayen, M, Grierson, D: Identification of a tomato gene for the ethylene-forming enzyme by expression in yeast. Proc Natl Acad Sci USA 88: 7434–7437 (1991).Google Scholar
  20. 20.
    Holdsworth, MJ, Bird, CR, Ray, J, Schuch, W, Grierson, D: Structure and expression of an ethylene-related mRNA from tomato. Nucl Acids Res 15: 731–739 (1987).Google Scholar
  21. 21.
    Holdsworth, MJ, Schuch, W, Grierson, D: Organization and expression of a wound/ripening-related small multigene family from tomato. Plant Mol Biol 11: 81–88 (1988).Google Scholar
  22. 22.
    Huang, P-L, Parks, JE, Rottman, WH, Theologis, A: Two genes encoding 1-aminocyclopropane-1-carboxylate synthase in zucchini (Cucurbita pepo) are clustered and similar but differentially regulated. Proc Natl Acad Sci USA 88: 7021–7025 (1991).Google Scholar
  23. 23.
    Hyodo, H: Stress/wound ethylene. In: Mattoo, AK, Suttle, JC (eds) The Plant Hormone Ethylene, pp. 43–63. CRC Press, Boca Raton, FL (1991).Google Scholar
  24. 24.
    Itzhaki, H, Woodson, WR: Characterization of an ethylene-responsive glutathione s-transferase gene cluster in carnation. Plant Mol Biol 22: 43–58 (1993).Google Scholar
  25. 25.
    Kende, H: Ethylene biosynthesis. Annu Rev Plant Physiol Plant Mol Biol 44: 283–307 (1993).Google Scholar
  26. 26.
    Köck, M, Hamilton, A, Grierson, D: eth1, a gene involved in ethylene synthesis in tomato. Plant Mol Biol 17: 141–142 (1991).Google Scholar
  27. 27.
    Liang, X, Abel, S, Keller, JA, Shen, NF, Theologis, A: The 1-aminocyclopropane-1-carboxylate synthase gene family of Arabidopsis thaliana. Proc Natl Acad Sci USA 89: 11046–11050 (1992).Google Scholar
  28. 28.
    Matsuda, J, Okabe, S, Hashimoto, T, Yamada, Y: Molecular cloning of hyoscyamine 6β-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger. J Biol Chem 266: 9460–9464 (1991).Google Scholar
  29. 29.
    McGarvey, DJ, Yu, H, Christoffersen, RE: Nucleotide sequence of a ripening-related cDNA from avocado fruit. Plant Mol Biol 15: 165–167 (1990).Google Scholar
  30. 30.
    McGarvey, DJ, Christoffersen, RE: Characterization and kinetic parameters of ethylene-forming enzyme from avocado fruit. J Biol Chem 267: 5964–5967 (1992).Google Scholar
  31. 31.
    Michael, MZ, Savin, KW, Baudinette, SC, Graham, MW, Chandler, SF, Lu, C-Y, Caesar, C, Gautrais, I, Young, R, Nugent, GD, Stevenson, KR, O'Connor, EL-J, Cobbett, CS, Cornish, EC: Cloning of ethylene biosynthetic genes involved in petal senescence of carnation and petunia, and their antisense expression in transgenic plants. In: Pech, JC, Latché, A, Balagué, C (eds) Cellular and Molecular Aspects of the Plant Hormone Ethylene, pp. 298–303. Kluwer Academic Publishers, Dordrecht, Netherlands (1993).Google Scholar
  32. 32.
    Montgomery, J, Goldman, S, Deikman, J, Margossian, L, Fischer, RL: Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc Natl Acad Sci USA 90: 5939–5943 (1993).Google Scholar
  33. 33.
    Mutschler, M, Guttieri, M, Kinzer, S, Grierson, D, Tucker, G: Changes in ripening-related processes in tomato conditioned by the alc mutant. Theor Appl Genet 76: 285–292 (1988).Google Scholar
  34. 34.
    Olson, DC, White, JA, Edelman, L, Harkins, RN, Kende, H: Differential expression of two genes for 1-aminocyclopropane-1-carboxylate synthase in tomato fruit. Proc Natl Acad Sci USA 88: 5340–5344 (1991).Google Scholar
  35. 35.
    Park, KY, Drory, A, Woodson, WR: Molecular cloning of an 1-aminocyclopropane-1-carboxylate synthase from senescing carnation flower petals. Plant Mol Biol 18: 377–386 (1992).Google Scholar
  36. 36.
    Peck, SC, Olson, DC, Kende, H: A cDNA sequence encoding 1-aminocyclopropane-1-carboxylate oxidase from pea. Plant Physiol 101: 689–690 (1993).Google Scholar
  37. 37.
    Reid, MS, Wu, M-J: Ethylene in flower development and senescence. In: Mattoo, AK, Suttle, JC (eds) The Plant Hormone Ethylene, pp. 215–234. CRC Press, Boca Raton, FL (1991).Google Scholar
  38. 38.
    Rottmann, WH, Peter, GF, Oeller, PW, Keller, JA, Shen, NF, Nagy, BP, Taylor, LP, Campbell, AD, Theologis, A: 1-aminocyclopropane-1-carboxylate synthase in tomato is encoded by a multigene family whose transcription is induced during fruit and floral senescence. J Mol Biol 222: 937–961 (1991).Google Scholar
  39. 39.
    Sanger, F, Nicklen, SC, Coulsen, AR: DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  40. 40.
    Spanu, R, Reinhardt, D, Boller, T: Analysis and cloning of the ethylene-forming enzyme from tomato by functional expression of its mRNA in Xenopus laevis oocytes. EMBO J 10: 2007–2013 (1991).Google Scholar
  41. 41.
    Ververidis, P, John, P: Complete recovery in vitro of ethylene-forming enzyme activity. Phytochem 30: 725–727 (1991).Google Scholar
  42. 42.
    Wang, H, Woodson, WR: Reversible inhibition of ethylene action and interruption of petal senescence in carnation flowers by norbornadiene. Plant Physiol 89: 434–438 (1989).Google Scholar
  43. 43.
    Wang, H, Woodson, WR: A flower senescence-related mRNA from carnation shares sequence similarity with fruit ripening-related mRNAs involved in ethylene biosynthesis. Plant Physiol 96: 1000–1001 (1991).Google Scholar
  44. 44.
    Wang, H, Woodson, WR: Nucleotide sequence of a cDNA encoding the ethylene-forming enzyme from petunia corollas. Plant Physiol 100: 535–536 (1992).Google Scholar
  45. 45.
    Woodson, WR, Lawton, KA: Ethylene-induced gene expression in carnation petals. Relationship to autocatalytic ethylene production and senescence. Plant Physiol 87: 498–503 (1988).Google Scholar
  46. 46.
    Woodson, WR, Park, KY, Drory, A, Larsen, PB, Wang, H: Expression of ethylene biosynthetic pathway transcripts in senescing carnation flowers. Plant Physiol 99: 526–532 (1992).Google Scholar
  47. 47.
    Zarembinski, TI, Theologis, A: Anaerobiosis and plant growth hormones induce two genes encoding 1-aminocyclopropane-1-carboxylate synthase in rice (Oryza sativa L.). Mol Biol Cell 4: 363–373 (1993).Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Xiaoyan Tang
    • 1
  • Hong Wang
    • 1
  • Amanda S. Brandt
    • 1
  • William R. Woodson
    • 1
  1. 1.Department of HorticulturePurdue UniversityWest LafayetteUSA

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